A photoaffinity ligand of the acetylcholine‐binding site predominantly labels the region 179–207 of the α‐subunit on native acetylcholine receptor from Torpedo marmorata

FEBS Letters

Giraudat, Jérôme¹  Hirth, Christian¹  Chang, Jui-Yoa¹  Goeldner, Maurice¹  Kotzyba-Hibert, Florence¹  Dennis, Michael¹  Changeux, Jean-Pierre¹ 

[1] Unité de Neurobiologie Moléculaire, Unité Associée au Centre National de la Recherche Scientifique, UA 041149, Interactions Moléculaires et Cellulaires, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cédex 15, France

关键词: Acetylcholine receptor;    Photoaffinity labeling;    Protein structure;    Acetylcholine-binding site;    Aryldiazonium salt;    HPLC;    high-performance liquid chromatography;    TFA;    trifluoroacetic acid;    HFBA;    heptafluorobutyric acid;   

DOI  :  10.1016/0014-5793(86)81497-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Regions of the Torpedo marmorata acetylcholine receptor (AChR) α-subunit involved in the binding of acetylcholine were probed with two different covalent ligands. The sulfhydryl-directed affinity reagent 4-(N-maleimido)phenyltrimethylammonium iodide labeled a single α-subunit cyanogen bromide fragment on the reduced AChR which was identified as α 179–207. The novel photoaffinity ligand p-(N,N-dimethylamino)-benzenediazonium fluoroborate, on the other hand, labeled three distinct α-chain cyanogen bromide fragments on the unmodified AChR in a carbamylcholine-protectable manner. The major radiolabeled species was purified and identified by sequence analysis as α 179–207. The acetylcholine-binding site on the native AChR may thus involve several distinct portions of the α-chain, with the region α 179–207 making a major contribution to the site.

【 授权许可】

Unknown